Methods of removing alpha-galactose

Abstract

Tissue products lacking a desired percentage of immunogenic epitopes, such as galactose alpha-1,3 galactose epitopes, are provided. Methods of making and using the tissue products are also provided.

What is claimed is: 
     
       1. A method of preparing a dermal tissue product, comprising:
 selecting at least one collagen-containing tissue matrix contained within a cellular dermal tissue and containing galactose alpha-1,3-galactose moieties; and 
 contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase under conditions sufficient to remove galactose alpha-1,3-galactose moieties from the tissue matrix contained within the cellular dermal tissue; 
 thereby preparing a dermal tissue product. 
 
     
     
       2. The method of  claim 1 , wherein the method further comprises decellularizing the cellular dermal tissue following contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase. 
     
     
       3. The method of  claim 2 , wherein the tissue matrix is completely decellularized. 
     
     
       4. The method of  claim 1 , wherein galactose alpha-1,3-galactose is substantially or completely removed from the collagen-containing tissue matrix. 
     
     
       5. The method of  claim 1 , wherein the alcalase removes galactose alpha-1,3-galactose moieties without damaging the at least one tissue matrix. 
     
     
       6. The method of  claim 1 , further comprising performing an assay to determine if galactose alpha-1,3-galactose moieties have been removed from the at least one collagen-containing tissue matrix. 
     
     
       7. The method of  claim 6 , wherein the assay comprises measuring a concentration of galactose alpha-1,3-galactose moieties in the collagen-containing tissue matrix. 
     
     
       8. The method of  claim 6 , wherein the assay comprises an histochemical assay. 
     
     
       9. The method of  claim 6 , wherein the assay comprises an immunoassay. 
     
     
       10. The method of  claim 1 , wherein the alcalase is used at a concentration ranging from about 0.0001% to about 0.1% and for a period of time ranging from about 0.5 hours to about 24 hours. 
     
     
       11. The method of  claim 1 , further comprising contacting the tissue with alpha-galactosidase following contacting the at least one collagen-containing tissue matrix contained within the cellular dermal tissue with alcalase. 
     
     
       12. The method of  claim 1 , wherein tissue from one or more different animals are used. 
     
     
       13. The method of  claim 1 , wherein the at least one collagen-containing tissue matrix is a porcine tissue matrix. 
     
     
       14. The method of  claim 2 , further comprising adding one or more viable and histocompatible cells to the tissue product. 
     
     
       15. The method of  claim 14 , wherein the one or more cells are mammalian cells. 
     
     
       16. The method of  claim 14 , wherein the one or more cells are stem cells. 
     
     
       17. The method of  claim 1 , further comprising treating the at least one collagen-containing tissue matrix to reduce bioburden. 
     
     
       18. The method of  claim 17 , wherein treating the at least one collagen-containing tissue matrix to reduce bioburden comprises irradiating the tissue product.